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From 2010.igem.org

Contents 1 136. labday 01.10.2010 1.1 mini preps of CD clones 1.2 test digestion of CD clones 1.3 Biobrick assembly pSB1C3_lITR_hTERT_beta-globin_CD

136. labday 01.10.2010

mini preps of CD clones

Investigator: Kira

c(p689)= 115 ng/ul
c(690)= 151, 20 ng/ul
c(691)= 122,27 ng/ul
c(p692) = 126,42 ng/ul

test digestion of CD clones

Investigator: Kira

Components	sample Volume/µL
DNA	4,0 µl
BSA (10x)	2 µl
Buffer no. 4	2,0 µl
Enzyme 1 XbaI	1,0 µl
Enzyme 2 AgeI	1,5 µl
H2O	9,5 µl
Total volume	20

incubation @ 37 C for approx. 2 h

1% agarose gel

Biobrick assembly pSB1C3_lITR_hTERT_beta-globin_CD

Investigator: Kira

c(pSB1C3_lITR_hTERT_beta-globin)= 333 ng/ul
c(pSB1C3_CD)= 151 ng/ul

Components	vector Volume/µL	insert Volume/µL
DNA	4,5 µl	6
BSA (10x)	3 µl	3
Buffer no. 4	3,0 µl	3
Enzyme 1 XbaI	0 µl	1,5
Enzyme 2 SpeI	1,5 µl	0
Enzyme 3 PstI-HF	1,0	1
H2O	17	15,5
Total volume	25

incubation @ 37 C for approx. 2 h

1% agarose gel

Ligation

DNA-mix: 8 ul (vector..)+(insert...)
T4 ligase: 1 ul
T4 buffer: 1 ul
Incubation @ RT for 30 min

Transformation was performed according to the standard protocol w BL21 cells.